Re: Hematoxylin & Eosin Staining on Frozen Brain

From:

"J. A. Kiernan"

Erica.Schaffer@aventis.com wrote:
> I am trying to find an H&E protocol for staining frozen brain sections cut at
> 7 microns. I have tried to stain the samples using the same protocol I use
> for other tissue types but I am not getting good results. If anyone has any
> advice, I would really appreciate it.
If the frozen sections are of unfixed tissue you should
fix them before staining. Alcohol or an alcohol-acetic
mixture for a few minutes should be OK.
If you really want your frozen sections to look like
paraffin sections of formaldehyde fixed tissue, immerse
in formaldehyde for 12-48 hours, wash, dehydrate in
100% alcohol, and rehydrate before staining.
After thorough formaldehyde fixation cell nuclei
look less interesting than after a coagulant fixative,
which generates characteristic chromatin patterns in
different cell types. (The boring formaldehyde nuclei
probably more closely resemble the pre-death cellular
architecture!) Eosin staining is weaker after adequate
formaldehyde fixation than after a simple coagulant
fixative, but it's easy to adjust this counterstain.
With regard to brain, it's important to know that
_H&E is a lousy stain for the CNS_ even after ideal
fixation etc. There are lots of better methods, and
many of them are easier to do than H&E.
--
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada N6A 5C1
kiernan@uwo.ca
http://publish.uwo.ca/~jkiernan/